Lasmiditan promotes recovery from acute kidney injury through induction of mitochondrial biogenesis

Abstract

Acute kidney injury (AKI) involves rapid loss of renal function and occurs in 8-16% of hospitalized patients. AKI can be induced by drugs, sepsis, and ischemia-reperfusion (I/R). Hallmarks of AKI include mitochondrial and microvasculature dysfunction as well as renal tubular injury. There is currently no available therapeutic for AKI. Previously, our group identified that serotonin (5-HT)_1F receptor agonism with lasmiditan accelerated endothelial cell recovery and induced mitochondrial biogenesis (MB) in vitro. We hypothesized that lasmiditan, a Federal Drug Administration-approved drug, would induce MB and improve microvascular and renal function in a mouse model of AKI. Male mice were subjected to renal I/R and treated with lasmiditan (0.3 mg/kg) or vehicle beginning 24 h after injury and then daily until euthanasia at 6 or 12 days. Serum creatinine was measured to estimate glomerular filtration rate. The renal cortex was assessed for mitochondrial density, vascular permeability and integrity, tubular damage, and interstitial fibrosis. Lasmiditan increased mitochondrial number (1.4-fold) in renal cortices. At 6 days, serum creatinine decreased 41% in the I/R group and 72% with lasmiditan. At 6 or 12 days, kidney injury molecule-1 increased in the I/R group and decreased 50% with lasmiditan. At 12 days, interstitial fibrosis decreased with lasmiditan by 50% and collagen type 1 by 38%. Evan's blue dye leakage increased 2.5-fold in the I/R group and was restored with lasmiditan. The tight junction proteins zonula occludens-1, claudin-2, and claudin-5 decreased in the I/R group and recovered with lasmiditan. At 6 or 12 days, peroxisome proliferator-activated receptor-γ coactivator-1α and electron transport chain complexes increased only with lasmiditan. In conclusion, lasmiditan treatment beginning AKI induces MB, attenuated vascular and tubular injury, decreased interstitial fibrosis, and lowered serum creatinine. Given that lasmiditan is a Federal Drug Administration-approved drug, these preclinical data support repurposing lasmiditan as a therapeutic for AKI.NEW & NOTEWORTHY AKI pathology involves a rapid decline in kidney function and occurs in 8-16% of hospitalized patients. There is currently no therapeutic for AKI. AKI results in mitochondria dysfunction, microvasculature injury, and loss of renal tubular function. In an I/R-induced AKI mouse model, treatment with the FDA-approved 5-HT_1F receptor-selective agonist lasmiditan induced mitochondrial biogenesis, improved vascular integrity, reduced fibrosis, and reduced proximal tubule damage. These data support repurposing lasmiditan for the treatment of AKI.

Keywords: acute kidney injury; fibrosis; microvasculature injury; mitochondrial biogenesis; mitochondrial dysfunction.

Graphical abstract

Figure 1.

Mitochondrial number increases in the mouse kidney cortex after lasmiditan treatment in vivo. Mice were treated with vehicle (V) or 0.3 mg/kg lasmiditan (L) with two doses (0 and 24 h) and harvested after 2 days. The kidney cortex was harvested, processed, and imaged via transmission electron microscopy. A: representative photomicrographs of the kidney cortex. B: mitochondrial number/field of five cells in each treatment group were enumerated. C: mitochondrial area averages of each group. Data are expressed as means ± SE; n = 6. P < 0.05 by a two-tailed unpaired t test. ^a,bDifferent letters on the top of the bars represent different statistical significance.

Figure 2.

Treatment with lasmiditan (L) increases recovery of renal function and decreases vascular permeability after ischemia-reperfusion (I/R) injury. A: serum creatinine was assessed 1 and 6 or 12 days after renal I/R injury. B: histopathology analysis of tubular damage at 1 and 12 days. C: hematoxylin and eosin staining of whole kidney slides. The black arrows illustrate dilated tubules. The red arrows illustrate dilated tubules that are filled with eosinophilic material (casts). D and E: immunoblot analysis of kidney injury molecule-1 (Kim-1) expression was used to assess tubular injury at 6 days (D) or 12 days (E). F: vascular leakage was assessed using Evan’s blue dye (EBD). G−I: immunoblot analysis of zonula occludens-1 (ZO-1; G), claudin-5 (H), and claudin-2 (I). J: representative immunoblot images. Data are expressed as means ± SE; n = 6–8. P < 0.05 by a Shapiro-Wilk test and nonparametric ANOVA (Kruskal–Wallis H test followed by a Dunnet’s post hoc test). ^a,b,cDifferent letters on the top of the bars represent different statistical significance. V, vehicle.

Figure 3.

Treatment with lasmiditan (L) restores peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) and mitochondrial complexes after ischemia-reperfusion (I/R) injury. A: immunoblot analysis of PGC-1α at 6 days. Subunits of complex V (B) and complex IV (C) at 6 days. D: a cocktail of five antibodies was used to assess changes in mitochondrial complexes at 6 days. E: immunoblot analysis of PGC-1α at 12 days. F and G: subunits of complex IV (F) and complex V (G) at 12 days. H: immunoblot of the antibody cocktail at 12 days. Data are expressed as means ± SE; n = 6–8. P < 0.05 by a Shapiro–Wilk test and one-way ANOVA (followed by a Tukey’s post hoc test). ^a,bDifferent letters on the top of the bars represent different statistical significance. V, vehicle.

Figure 4.

Treatment with lasmiditan (L) reduces tubulointerstitial fibrosis and collagen type 1 after ischemia-reperfusion (I/R) injury. A: immunoblot analysis of collagen type 1 expression at 12 days. B: tubulointerstitial fibrosis scoring from fibrotic kidney cortex tissue at 1 and 12 days after I/R injury. C: Masson’s trichrome-stained representative images for fibrosis scoring. The yellow arrows illustrate blue-stained collagen fibers (fibrosis), which are minimally increased in a region with tubular regeneration and interstitial inflammation. Note that the cast material is stained blue. Data are expressed as means ± SE; n = 6–8. P < 0.05 by a Shapiro–Wilk test and nonparametric ANOVA (Kruskals–Wallis H test followed by a Dunnet’s post hoc test). ^a,bDifferent letters on the top of the bars represent different statistical significance.